A simple and direct pre-embedding technique for ultrastructure of scarce biological specimens.
A simple and rapid technique for pre-embedding scarce biological specimens for Transmission electron microscopy (TEM) is reported. It is based on pre-embedding biological samples in bovine serum albumin (BSA) and bis-acrylamide (BA), cross-linked and polymerized with paraformaldehyde, glutaraldehyde, ammonium persulfate and Temed. Pre-embedding in BSA and BA offers several advantages over traditional pre-embedding techniques for TEM including the ability to visualize the sample and a more resistant matrix. This results in more reproducible and consistent analysis. It can be applied to tissues, cells, and subcellular structures handled as pellets or suspensions. In addition, use of the pre-embedding matrix for light microscopy is reported. The ability to pre-embed scarce biological specimens efficiently and reproducibly provides a valuable way to study and characterize cytological tissues such as biopsies or cystic and amniotic fluid cells.
Taupin
P
P
Fighting Blindness Vision Research Institute, National Institute for Cellular Biotechnology and Dublin City University, Dublin 9, Glasnevin, Ireland. philippe.taupin@dcu.ie
D016475:3T3 Cells; D000645:Ammonium Sulfate / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000378:metabolism; D005029:Ethylenediamines / Q000378:metabolism; D005557:Formaldehyde / Q000378:metabolism; D005976:Glutaral / Q000378:metabolism; D006652:Histological Techniques / Q000379:methods*; D006801:Humans; D007668:Kidney / Q000166:cytology; D051379:Mice; D008853:Microscopy; D046529:Microscopy, Electron, Transmission; D011108:Polymers / Q000378:metabolism; D015203:Reproducibility of Results; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013048:Specimen Handling / Q000379:methods; D016610:Tissue Embedding / Q000379:methods*
C106521:BANG polymer gel; D003432:Cross-Linking Reagents; D005029:Ethylenediamines; D011108:Polymers; C005798:N,N,N',N'-tetramethylethylenediamine; D005557:Formaldehyde; C031276:ammonium peroxydisulfate; D012710:Serum Albumin, Bovine; D000645:Ammonium Sulfate; D005976:Glutaral; C003043:paraform